Figure 1 | Scientific Reports

Figure 1

From: Doublecortin-like kinase 1 promotes hepatocyte clonogenicity and oncogenic programming via non-canonical β-catenin-dependent mechanism

Figure 1

DCLK1-expressing primary human hepatocytes form spheroids in 3D levitated culture devoid of matrigel. (a) Monolayer cultures of primary human hepatocytes in complete hepatocyte media were infected with lentiviruses expressing GFP (control) or GFP-tagged human DCLK1 for 48 h. Ten thousand hepatocytes from each trypsinized culture were used for magnetic levitation assay in 6-well ultra-low attachment plates. On day 5, live cell imaging was carried out to record spheroids formation (red arrows, magnification × 10, upper panel). The levitated culture of hepatocytes transduced with lentiviruses expressing RFP (control) is shown in lower panel. (b) Live cell images of the spheroids (magnification × 40) in bright field (upper panel)) and for GFP expression (green, lower panel) to show that the spheroids were developed from GFP-DCLK1-expressing primary human hepatocytes. (c) GS5 cells derived from Huh7.5 hepatoma cell line and harbor an HCV subgenomic replicon expressing GFP-NS5A were used as a positive control under similar levitation assay conditions.

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